Secretory Component antibody (Membrane-Bound, Soluble)

Catalog No. ABIN457936

Product Details anti-Secretory Component Antibody

Target: Secretory Component

Binding Specificity: Membrane-Bound, Soluble

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: This Secretory Component antibody is un-conjugated

Application: ELISA, Western Blotting (WB), Immunofluorescence (IF), Haemagglutination (H)

Specificity: The antiserum does not react with any other component of the human Ig system or any other human plasma protein as tested. This antiserum has not been tested for cross-reactivity with other species.

Characteristics: Monoclonal mouse antiserum to human secretory component, free and bound

Immunogen: Highly purified secretory component isolated from human milk.

Clone: NI 194-4 (A89-039)

Isotype: IgG1 kappa

Application Details

Application Notes: To identify the presence of free or bound secretory component in human serum, other body fluids, cell and tissue substrates and to determine its concentration in techniques as radio immuno assay, ELISA, indirect immunoperoxidase and immunofluorescence staining, haemagglutination and immunoblotting. The optimum working dilution is an assay-related characteristic and should always be determined by titration. For histochemical use optimum dilutions are mostly from 1:50 to 1:200, in ELISA from 1:500 upwards, in Western blotting from 1:1000 upwards. These data should be interpreted as general recommendations only.

Restrictions: For Research Use only

Independent Validation (WB)

Validation #103267 (Western Blotting)

by: Secretory IgA, Inc.

No.: #103267

Date: 06/05/2019

Antigen: Secretary Component

Lot Number: 4628

Method validated: Western Blotting

Positive Control:

Recombinant human secretory component

Negative Control:

Purified plasma Human immunoglobulin A without added secretory component

Notes:

Passed. ABIN457936 specifically recognizes human secretory Component free or conjugated to dimeric or polymeric human IgA.

Validation Images

Full Methods

Primary Antibody: ABIN457936

Secondary Antibody: sheep anti-mouse HRP (Amersham, NA931V, lot 16964881)

Full Protocol:

• Conjugate recombinant human Secretory Component (rhSC) to Jacalin-purified human plasma immunoglobulin A (IgA).
• Determine total protein content of the protein solution at OD₂₈₀.
• Denature 10µg of total protein for 5min at 95°C in 12.5µL of NuPAGE LDS buffer sample (Invitrogen, NP0007) in a total volume of 50µL and subsequently separate them on a NuPAGE Tris-Acetate 4-8% gel (Invitrogen, EA0375) for 75min at 150V.
• Transfer proteins onto membrane (Invitrogen, IB301001) with an iBlot Western blotting system for 3.5h at 300mA.
• Block the membrane in TBST (Boston Bioproduct, IBB-180, lot C15K132) and 5% blotting-grade blocker (invitrogen, 1706404, lot L005668A), for 1h at RT under low agitation.
• Incubation with primary mouse anti-Secretory Component antibody (antibodies-online, ABIN457936, lot 4628) diluted 1:1500 in TBST containing 5% blocker ON at 4°C under slow agitation.
• Wash membrane 6x for 5min with TBST.
• Incubation with secondary sheep anti-mouse HRP (Amersham, NA931V, lot 16964881) diluted 1:4000in TBS-tween for 1h at RT.
• Wash membrane 6x for 5min with TBST.
• Reveal protein bands using ECL prime western blotting Detection reagent(Sigma-Aldrich, RPN2236, lot 16961638) on a FluorChem E system (protein simple, SN FE0238).

Experimental Notes:

The human Secretory Component antibody ABIN457936 reveals a protein of the expected molecular weight of unconjugated human Secretory Component in the purified lane and the high molecular weight complex when conjugated with plasma IgA. The protein bands is only visible in the positive but not the negative controls.

Handling

Format: Lyophilized

Concentration: IgG concentration is 1 mg/ml. No foreign proteins added.

Buffer: Delipidated, heat inactivated, lyophilized, stable whole ascites

Storage: 4 °C/-20 °C

Storage Comment: The lyophilized product is shipped at ambient temperature and may be stored at +4 °C, prolonged storage at or below -20 °C. Reconstitute the lyophilized ascites by adding 0.5 ml sterile distilled water. Dilutions may be prepared by adding phosphate buffered saline (PBS, pH 7.2). Avoid repeated thawing and freezing. If a slight precipitation occurs upon storage, this should be removed by centrifugation and will not affect the performance of the product. Diluted ascites should be stored at +4 °C, not refrozen, and preferably used the same day.

Target Details for Secretory Component

Target: Secretory Component

Abstract: Secretory Component Products

Synonyms: RNPIGR2 antibody, pIgA-R antibody, polymeric immunoglobulin receptor antibody, PIGR antibody, Pigr antibody

Background: The reactivity of the antiserum is restricted to determinants on the secretory component as tested in haemagglutination, haemagglutination inhibition, indirect binding enzyme immunoassay, competitive inhibition enzyme immunoassay, immunoblotting, immunoprecipitation, latex agglu-tination assay and histochemistry (Results of an IUIS/WHO collaborative study, Mestecky J. et al. (1996) J. Immunol. Methods 193, 103-148).